The Patterning and Proportion of Charged Residues in the Arginine-Rich Mixed-Charge Domain Determine the Membrane-Less Organelle Targeted by the Protein
Abstract
:1. Introduction
2. Results
2.1. Charge Patterning in the R-MCDs Influences Their Distribution to MLOs
2.2. Charge Patterning Affects the Interactome of R-MCDs
2.3. A Net Positive Charge in R-MCDs Enhances Their Incorporation into MLOs
2.4. Charge Patterning in Natural R-MCDs Determines Protein Distribution to Specific MLOs
2.5. Charge Patterning Determines the Phase-Separating Properties of R-MCD Peptides
3. Discussion
4. Materials and Methods
4.1. Recombinant Plasmids Construction
4.2. Transfection of HeLa Cells and Image Acquisition by Confocal Microscopy
4.3. Peptide Synthesis and Purification of Recombinant mScarletI-NPM1
4.4. Phase Separation of Peptides
4.5. Proximity Labeling and Quantitative LC–MS
4.6. Colocalization Analysis
4.7. Bioinformatic Analysis of Biochemical Properties of Peptides and Proteins
4.8. Statistic Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Miyagi, T.; Yamazaki, R.; Ueda, K.; Narumi, S.; Hayamizu, Y.; Uji-i, H.; Kuroda, M.; Kanekura, K. The Patterning and Proportion of Charged Residues in the Arginine-Rich Mixed-Charge Domain Determine the Membrane-Less Organelle Targeted by the Protein. Int. J. Mol. Sci. 2022, 23, 7658. https://doi.org/10.3390/ijms23147658
Miyagi T, Yamazaki R, Ueda K, Narumi S, Hayamizu Y, Uji-i H, Kuroda M, Kanekura K. The Patterning and Proportion of Charged Residues in the Arginine-Rich Mixed-Charge Domain Determine the Membrane-Less Organelle Targeted by the Protein. International Journal of Molecular Sciences. 2022; 23(14):7658. https://doi.org/10.3390/ijms23147658
Chicago/Turabian StyleMiyagi, Tamami, Rio Yamazaki, Koji Ueda, Satoshi Narumi, Yuhei Hayamizu, Hiroshi Uji-i, Masahiko Kuroda, and Kohsuke Kanekura. 2022. "The Patterning and Proportion of Charged Residues in the Arginine-Rich Mixed-Charge Domain Determine the Membrane-Less Organelle Targeted by the Protein" International Journal of Molecular Sciences 23, no. 14: 7658. https://doi.org/10.3390/ijms23147658